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Transduction of human CD34+ CD38- bone marrow and cord blood-derived SCID-repopulating cells with third-generation lentiviral vectors.
- Source :
-
Molecular therapy : the journal of the American Society of Gene Therapy [Mol Ther] 2000 Jun; Vol. 1 (6), pp. 566-73. - Publication Year :
- 2000
-
Abstract
- The major limitations of Moloney murine leukemia virus (MoMLV)-based vectors for human stem cell applications, particularly those requiring bone marrow (BM) stem cells, include their requirement for mitosis and retroviral receptor expression. New vectors based upon lentiviruses such as HIV-1 exhibit properties that may circumvent these problems. We report that novel third-generation, self-inactivating lentiviral vectors, expressing enhanced green fluorescent protein (EGFP) and pseudotyped with vesicular stomatitis virus G glycoprotein (VSV-G), can efficiently transduce primitive human repopulating cells derived from human BM and cord blood (CB) tested by the SCID-repopulating cell (SRC) assay. Highly purified CD34+ CD38- CB or BM cells were efficiently transduced (4-69%) and stably expressed in EGFP for 40 days in culture following infection for only 24 h without fibronectin, polybrene, or cytokines. Nonobese diabetic/severe combined immune-deficient (NOD/SCID) mice transplanted with transduced cells from either CB or BM donors were well engrafted, demonstrating maintenance of SRC during the infection procedure. Serially obtained femoral BM samples indicated that the proportion of EGFP+ cells within both myeloid and lymphoid lineages was maintained or even increased over time, averaging 42.3 +/- 6.6% for BM donors and 23.3 +/- 7.2% for CB at 12 weeks. Thus, the third-generation lentivectors readily transduce human CB and BM stem cells, under minimal conditions of ex vivo culture, where MoMLV-based vectors are ineffective. Since CB is inappropriate for most therapeutic applications, the efficient maintenance and transduction of BM-derived SRC during the short infection procedure are notable advantages of lentivectors.
- Subjects :
- ADP-ribosyl Cyclase
ADP-ribosyl Cyclase 1
Animals
Antigens, CD34 metabolism
Antigens, Differentiation metabolism
Fetal Blood immunology
Gene Expression
Green Fluorescent Proteins
Hematopoietic Stem Cell Transplantation
Humans
Luminescent Proteins genetics
Membrane Glycoproteins
Mice
Mice, Inbred NOD
Mice, SCID
Moloney murine leukemia virus genetics
NAD+ Nucleosidase metabolism
Antigens, CD
Genetic Vectors
Hematopoietic Stem Cells immunology
Lentivirus genetics
Severe Combined Immunodeficiency immunology
Severe Combined Immunodeficiency therapy
Transduction, Genetic
Subjects
Details
- Language :
- English
- ISSN :
- 1525-0016
- Volume :
- 1
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Molecular therapy : the journal of the American Society of Gene Therapy
- Publication Type :
- Academic Journal
- Accession number :
- 10933981
- Full Text :
- https://doi.org/10.1006/mthe.2000.0077